In V itro Pollen Germ ina tion and Pollen Tube Growth of P inus thunbe rgii

LI Guo-ping; HUANG Qun-ce; YANG Lu-sheng; QIN Guang-yong

Volume 20 Issue 2

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In V itro Pollen Germ ina tion and Pollen Tube Growth of P inus thunbe rgii

1.
Provincial Key Laboratory of Ion Beam Bio-engineering, Zhengzhou University, Zhengzhou　450052, He’nan, China
2.
Department of Environment and Life Science, Putian University, Putian　351100, Fujian, China

Abstract

The organization of pollen and pollen tubes of Pinus thunbergiiwas studied by using a confocal laser scanning microscope after fluorescence labeling. A Pinus thunberg ii pollen was 4-celled pollen with two degenerated p rothallial cells, one generative cell and one tube cell. In culture, pollen germination occurred after app roximately 12h of hydration. During tube growth, the tube cell nucleusmoved into the elongating tube, but the generative cells remained within the pollen grain. There are two distinct zones in elongating pollen tubes. One began in the pollen andextended towards the pollen tube tip, containing abounding amylop lasts. The other zone was the clear zone lackingamylop lasts at the elongating pollen tube tip. The cytop lasmic-streaming type in the tube cellwas a regular fountainlike pattern, with organellesmoving towards the tip in the center of the tube and away from the tip along the cellcortex. Fluorescence microscopy indicated that the pollen tube wall contained cellulose microfibrils and, callose wasp resent in the younger pollen tubes, but disappeared from the older tubes. No callose p lug was detected within thepollen tube. The pollen tube generally developed two to six branches. The sucrose concentration in culture mediumaffected the formation of tube branching and the growth rate of tubes to a great extent. The p resent study suggestedthat the development and cellular organization of gymnosperms pollen tubeswere considerably different from those ofangiosperm pollen tubes.
- gymnosperms,
- Pinus thunbergii,
- pollen,
- pollen tube

References

[1]	Bedinger P. The remarkable biology of pollen[J]. Plant Cell, 1992,4 (8) : 879～887
[2]	[第一卷?修订本] [M]. 福州: 福建科学技术出版社, 1991: 282～292
[3]	Taylor L P, Hep ler P K. Pollen germination and tube growth [J].Annu Rev Plant Physiol PlantMol Biol, 1997, 48: 461～491
[4]	Carmichael J S, FriedmanW E. Double fertilization in Gnetum gnemon (Gnetaceae) : Its bearing on the evolution of sexual rep roduction within the Gnetales and the Anthophyte Clade[J]. Am J ofBot,1996, 83 (6) : 767～780
[5]	Derksen J. Pollen tubes: a model system for p lant cell growth [J].Bot Acta, 1996, 109: 341～345
[6]	Mascarenhas J P. Molecular mechanisms of pollen tube growth anddifferentiation[J]. Plant Cell, 1993, 5: 303～314
[7]	Monteiro D, Liu Q, Lisboa S, et al. Phosphoinositides and phosphatidic acid regulate pollen tube growth and reorientation through modulation of [Ca2 +] and membrane secretion [J]. J Exp Bot, 2005,56: 1665～1674
[8]	Singh H. Embryology of gymnosperms [A]. In: Zimmermann W,Carlquist S, Ozenda P, et al. Handbuch der Pflanzenanatomie: vol.10, part 2 [C]. Berlin: Gebrüder Borntraeger, 1978
[9]	林来官. 福建植物志
[10]	李国平,黄群策. 油杉( Keteleeria fortunei )花粉个体发育与传粉过程[J]. 林业科学, 2006, 42 (5) : 42～47
[11]	李国平,黄群策,秦广雍. 用激光扫描共聚焦显微镜观察雪松花粉和花粉管[J]. 激光生物学报, 2006, 15 (1) : 1～8
[12]	曹玉芳,许方,姚敦义. 松杉类植物雌雄配子体的发育与胚胎发生[J]. 莱阳农学院学报, 1995, 12 (3) : 206～212
[13]	吕世友. 云杉和油松花粉发育及其原叶细胞程序化死亡的研究[D]. 中国人民解放军军需大学, 2001: 31～48
[14]	deWin A H N, Knuiman B, Pierson E S, et al. Development andcellular organization of Pinus sylvestris pollen tubes[J]. Sex PlantRep rod, 1996, 9: 93～101
[15]	DawkinsM D, Owens J N. In vitro and in vivo pollen hydration,germination, and pollen tube growth in white sp ruce, Picea glauca(Moench) voss[J]. Int J Plant Sci, 1993, 164: 506～521
[16]	Anderson E D, Owens J N. Microsporogenesis, pollination, pollengermination and male gametophyte development in Taxus brevifolia[J]. Ann Bot, 2000, 86: 1033～1044
[17]	Fernando D D, Long SM, Sniezko R A. Growth and development ofconifer pollen tubes[J]. Sex Plant Rep rod, 2005, 18: 149～162
[18]	BarnabasB, FridvaiszkyL. Adhesion and germination of differentlytreated maize pollen grains on the stigma [J]. Acta Bot Hungar,1984, 30: 329～332
[19]	UwateW J, Lin J. Cytological zonation of Prunu savium L. pollentube in vivo[J]. J Ultrastruct Res, 1980, 71: 173～184
[20]	Justus C D, Anderhag P, Goins J L, et al. Microtubules and microfilaments coordinate to direct a fountain streaming pattern in elongating conifer pollen tube tip s [J]. Planta, 2004, 219: 103～109
[21]	CardenasL, Lovy-WheelerA, Wilsen KL, et al. Actin polymerization p romotes the reversal of streaming in the apex of pollen tubes[J]. CellMotil Cytoskeleton, 2005, 61: 112～127
[22]	Pierson E S, CrestiM. Cytoskeleton and cytop lasmic organization ofpollen and pollen tubes[J]. Int Rev of Cytol, 1992, 140: 73～125
[23]	Derksen J, Li Y Q, Knuiman B, et al. The wall of Pinus sylvestrisL. pollen tubes[J]. Protop lasma, 1999, 208: 26～36
[24]	LazzaroM D, Donohue JM, Soodavar FM. Disrup tion of cellulosesynthesis by isoxaben causes tip swelling and disorganizes corticalmicrotubules in elongating conifer pollen tubes [J]. Protop lasma,2003, 220: 201～207
[25]	Vervaeke I,Londers E, Piot G, et al. The division of the generativenucleus and the formation of callose p lugs in pollen tubes of Aechm ea fasciata (Bromeliaceae) cultured in vitro [J]. Sex Plant Rep rod, 2005, 18 (1) : 9～19
[26]	邢树平. 花粉管生长调控的研究进展[J]. 广西植物, 1998, 18(1) : 82～88
[27]	Pierson E S, Li YQ, Zhang H Q, et al. Pulsatory growth of pollentubes: investigation of a possible relationship with the periodic distribution of cell wall components [J]. Acta Bot Neerl, 1995, 44(2) : 121～128

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通讯作者: 陈斌, bchen63@163.com

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沈阳化工大学材料科学与工程学院沈阳 110142

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In V itro Pollen Germ ina tion and Pollen Tube Growth of P inus thunbe rgii

1. Provincial Key Laboratory of Ion Beam Bio-engineering, Zhengzhou University, Zhengzhou　450052, He’nan, China

2. Department of Environment and Life Science, Putian University, Putian　351100, Fujian, China

Abstract: The organization of pollen and pollen tubes of Pinus thunbergiiwas studied by using a confocal laser scanning microscope after fluorescence labeling. A Pinus thunberg ii pollen was 4-celled pollen with two degenerated p rothallial cells, one generative cell and one tube cell. In culture, pollen germination occurred after app roximately 12h of hydration. During tube growth, the tube cell nucleusmoved into the elongating tube, but the generative cells remained within the pollen grain. There are two distinct zones in elongating pollen tubes. One began in the pollen andextended towards the pollen tube tip, containing abounding amylop lasts. The other zone was the clear zone lackingamylop lasts at the elongating pollen tube tip. The cytop lasmic-streaming type in the tube cellwas a regular fountainlike pattern, with organellesmoving towards the tip in the center of the tube and away from the tip along the cellcortex. Fluorescence microscopy indicated that the pollen tube wall contained cellulose microfibrils and, callose wasp resent in the younger pollen tubes, but disappeared from the older tubes. No callose p lug was detected within thepollen tube. The pollen tube generally developed two to six branches. The sucrose concentration in culture mediumaffected the formation of tube branching and the growth rate of tubes to a great extent. The p resent study suggestedthat the development and cellular organization of gymnosperms pollen tubeswere considerably different from those ofangiosperm pollen tubes.

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Reference (27)

[1]	Bedinger P. The remarkable biology of pollen[J]. Plant Cell, 1992,4 (8) : 879～887
[2]	[第一卷?修订本] [M]. 福州: 福建科学技术出版社, 1991: 282～292
[3]	Taylor L P, Hep ler P K. Pollen germination and tube growth [J].Annu Rev Plant Physiol PlantMol Biol, 1997, 48: 461～491
[4]	Carmichael J S, FriedmanW E. Double fertilization in Gnetum gnemon (Gnetaceae) : Its bearing on the evolution of sexual rep roduction within the Gnetales and the Anthophyte Clade[J]. Am J ofBot,1996, 83 (6) : 767～780
[5]	Derksen J. Pollen tubes: a model system for p lant cell growth [J].Bot Acta, 1996, 109: 341～345
[6]	Mascarenhas J P. Molecular mechanisms of pollen tube growth anddifferentiation[J]. Plant Cell, 1993, 5: 303～314
[7]	Monteiro D, Liu Q, Lisboa S, et al. Phosphoinositides and phosphatidic acid regulate pollen tube growth and reorientation through modulation of [Ca2 +] and membrane secretion [J]. J Exp Bot, 2005,56: 1665～1674
[8]	Singh H. Embryology of gymnosperms [A]. In: Zimmermann W,Carlquist S, Ozenda P, et al. Handbuch der Pflanzenanatomie: vol.10, part 2 [C]. Berlin: Gebrüder Borntraeger, 1978
[9]	林来官. 福建植物志
[10]	李国平,黄群策. 油杉( Keteleeria fortunei )花粉个体发育与传粉过程[J]. 林业科学, 2006, 42 (5) : 42～47
[11]	李国平,黄群策,秦广雍. 用激光扫描共聚焦显微镜观察雪松花粉和花粉管[J]. 激光生物学报, 2006, 15 (1) : 1～8
[12]	曹玉芳,许方,姚敦义. 松杉类植物雌雄配子体的发育与胚胎发生[J]. 莱阳农学院学报, 1995, 12 (3) : 206～212
[13]	吕世友. 云杉和油松花粉发育及其原叶细胞程序化死亡的研究[D]. 中国人民解放军军需大学, 2001: 31～48
[14]	deWin A H N, Knuiman B, Pierson E S, et al. Development andcellular organization of Pinus sylvestris pollen tubes[J]. Sex PlantRep rod, 1996, 9: 93～101
[15]	DawkinsM D, Owens J N. In vitro and in vivo pollen hydration,germination, and pollen tube growth in white sp ruce, Picea glauca(Moench) voss[J]. Int J Plant Sci, 1993, 164: 506～521
[16]	Anderson E D, Owens J N. Microsporogenesis, pollination, pollengermination and male gametophyte development in Taxus brevifolia[J]. Ann Bot, 2000, 86: 1033～1044
[17]	Fernando D D, Long SM, Sniezko R A. Growth and development ofconifer pollen tubes[J]. Sex Plant Rep rod, 2005, 18: 149～162
[18]	BarnabasB, FridvaiszkyL. Adhesion and germination of differentlytreated maize pollen grains on the stigma [J]. Acta Bot Hungar,1984, 30: 329～332
[19]	UwateW J, Lin J. Cytological zonation of Prunu savium L. pollentube in vivo[J]. J Ultrastruct Res, 1980, 71: 173～184
[20]	Justus C D, Anderhag P, Goins J L, et al. Microtubules and microfilaments coordinate to direct a fountain streaming pattern in elongating conifer pollen tube tip s [J]. Planta, 2004, 219: 103～109
[21]	CardenasL, Lovy-WheelerA, Wilsen KL, et al. Actin polymerization p romotes the reversal of streaming in the apex of pollen tubes[J]. CellMotil Cytoskeleton, 2005, 61: 112～127
[22]	Pierson E S, CrestiM. Cytoskeleton and cytop lasmic organization ofpollen and pollen tubes[J]. Int Rev of Cytol, 1992, 140: 73～125
[23]	Derksen J, Li Y Q, Knuiman B, et al. The wall of Pinus sylvestrisL. pollen tubes[J]. Protop lasma, 1999, 208: 26～36
[24]	LazzaroM D, Donohue JM, Soodavar FM. Disrup tion of cellulosesynthesis by isoxaben causes tip swelling and disorganizes corticalmicrotubules in elongating conifer pollen tubes [J]. Protop lasma,2003, 220: 201～207
[25]	Vervaeke I,Londers E, Piot G, et al. The division of the generativenucleus and the formation of callose p lugs in pollen tubes of Aechm ea fasciata (Bromeliaceae) cultured in vitro [J]. Sex Plant Rep rod, 2005, 18 (1) : 9～19
[26]	邢树平. 花粉管生长调控的研究进展[J]. 广西植物, 1998, 18(1) : 82～88
[27]	Pierson E S, Li YQ, Zhang H Q, et al. Pulsatory growth of pollentubes: investigation of a possible relationship with the periodic distribution of cell wall components [J]. Acta Bot Neerl, 1995, 44(2) : 121～128

In V itro Pollen Germ ina tion and Pollen Tube Growth of P inus thunbe rgii

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In V itro Pollen Germ ina tion and Pollen Tube Growth of P inus thunbe rgii

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